2.1.- Cell lines and cell maintenance
The murine hybridoma cell line used in this work is the KB26.5 which produces an IgG3 antibody against antigen A1 of human red blood cells (used for ABO group determining in blood tests). KB26.5 cell line was transfected with BHRF1 (BamHI rightward reading frame 1)-enconding vector as described by Juanola et al. (2009).
Both cells lines were maintained in 125 mL polycarbonate shake flask (Corning Inc.) with a working volume of 20 mL incubated in a 5% CO2 air mixture and in a humidified atmosphere at 37ºC (Steri-cult 2000 Incubator). Flasks were continuously agitated at 110 rpm in an orbital shaking platform (Stuart SSL110). Cell passaging was performed every 2 or 3 days at an initial seeding density of 1.5·105 cells/mL.